Do high-nickel leaves shed by the nickel hyperaccumulator Alyssum murale inhibit seed germination of competing plants?

Elemental allelopathy suggests that nickel (Ni)-rich leaves shed by hyperaccumulators inhibit the germination and growth of nearby plant species. Here, the germination of eight herbaceous species following addition of Alyssum murale biomass or Ni(NO3)2, with the same Ni level added to soil, was assessed. The distribution of Ni in soil was tested by determining Ni phytoavailability and speciation over time. Phytoavailable Ni in soil amended with biomass declined rapidly over time due to Ni binding to iron (Fe)/manganese (Mn) oxides in the soil. No significant effects on seed germination were observed. Unlike the Ni complex in Alyssum biomass, more Ni remained soluble and phytoavailable in soil amended with Ni(NO3)2, thus significantly inhibiting seed germination. High-Ni leaves shed by hyperaccumulators did not appear to create a 'toxic zone' around the plants and inhibit germination or growth of competing plants. The lack of an allelopathic effect was probably related to low Ni availability.     

Phytoextraction potential of the nickel hyperaccumulators Leptoplax emarginata and Bornmuellera tymphaea

Leptoplax emarginata and Bornmuellera tymphaea are nickel hyperaccumulators of the Brassicaceae family endemic to serpentine soils in Greece. The aims of this work were to compare the growth and uptake behavior of these plants with the Ni hyperaccumulator species Thlaspi caerulescens and Alyssum murale, and to evaluate their effect on soil Ni availability. Plants were grown for 3 mo on three soils that differ in Ni availability. Ni availability in soils was measuredby isotopic exchange kinetics and DTPA-TEA extractions. Results showed that L. emarginata produced significantly more biomass than other plants. On the serpentine soil, B. tymphaea showed the highest Ni concentration in shoots. However, Niphytoextraction on the three soils was maximal with L. emarginata. The high initial Ni availability of soil Serp (470.5 mg kg(-1)) was the main explanation for the high Ni concentrations measured in plant shoots grown on this soil, compared to those grown on soils Calc and Silt A. murale was the least efficient in reducing Ni availability on the serpentine soil L. emarginata appeared as the most efficient species for Ni phytoextraction and decrease of the Ni available pool.     

Hyperaccumulator Alyssum murale relies on a different metal storage mechanism for cobalt than for nickel

The nickel (Ni) hyperaccumulator Alyssum murale has been developed as a commercial crop for phytoremediation/phytomining Ni from metal-enriched soils. Here, metal co-tolerance, accumulation and localization were investigated for A. murale exposed to metal co-contaminants. A. murale was irrigated with Ni-enriched nutrient solutions containing basal or elevated concentrations of cobalt (Co) or zinc (Zn). Metal localization and elemental associations were investigated in situ with synchrotron X-ray microfluorescence (SXRF) and computed-microtomography (CMT). A. murale hyperaccumulated Ni and Co (> 1000 microg g(-1) dry weight) from mixed-metal systems. Zinc was not hyperaccumulated. Elevated Co or Zn concentrations did not alter Ni accumulation or localization. SXRF images showed uniform Ni distribution in leaves and preferential localization of Co near leaf tips/margins. CMT images revealed that leaf epidermal tissue was enriched with Ni but devoid of Co, that Co was localized in the apoplasm of leaf ground tissue and that Co was sequestered on leaf surfaces near the tips/margins. Cobalt-rich mineral precipitate(s) form on leaves of Co-treated A. murale. Specialized biochemical processes linked with Ni (hyper)tolerance in A. murale do not confer (hyper)tolerance to Co. A. murale relies on a different metal storage mechanism for Co (exocellular sequestration) than for Ni (vacuolar sequestration).     

Effects of nickel hyperaccumulation on physiological characteristics of Alyssum murale grown on metal contaminated waste amended soil

A pot experiment was conducted to investigate the effect of nickel concentration on physiological characteristics of Alyssum murale when grown in a soil mixed with sewage sludge (at the rate of 2.8%). Two types of sludge were used: agricultural sewage sludge (S1) and industrial sewage sludge with an increasing nickel concentration (S2, S3, and S4). Results showed that Ni in shoots was higher than Ni in roots. A. murale is able to concentrate up to 12730 mg/kg Ni in leaves. The highest dry matter yield was observed with plants grown with agricultural sewage sludge. An addition of S2 and S3 increased shoot biomass. However, application of S4 reduced 40% shoot dry weight as compared to the control Ni treatment did not affect all chlorophyll fluorescence parameters. The F(v)/F(m) ratio was stable between Ni treatments. Photosynthesis rate (A) increased with agricultural sewage sludge, but remained stable with variable Ni rates from the industrial sludge. The chlorophyll content increased with S1, S2 and S3 but it remains constant with S4 when compared to the control Therefore, high nickel concentration did not affect the function of the photosynthetic machine of A. murale.     

接种泡囊假单胞菌对土壤生物性质及A. corsicum吸收Ni的影响

采用室内模拟试验方法,研究了在水稻土、元江土和墨江土中添加泡囊假单胞菌（Pseulormanas vesicularis）后土壤中微生物种群数量、土壤酶活性和镍超积累植物Alyssum corsicum对土壤镍的富集效果.土壤接种泡囊假单胞菌70d后,水稻土中DTPA提取态镍较对照土中的明显减少、元江土和墨江土中的有所减少;土壤中细菌、真菌和放线菌数量增加,5种土壤酶活性提高.试验结果表明,水稻土、元江土、墨江土添加泡囊假单菌后植物地上部生物量较对照分别增加了29%、309%和43%,进而提高了A.corsicum自土壤中富集镍的效率：水稻土中增加54%,元江土中增加306%,墨江土中增加32%.泡囊假单胞菌这一新用途的发现,可为植物修复微生物制剂和基因工程菌的开发提供本土的微生物的菌种资源.     

Exogenous cytokinin treatments of an Ni hyper-accumulator, Alyssum murale, grown in a serpentine soil: implications for phytoextraction

Application of exogenous plant growth regulators was examined as a viable technique to increase the efficiency of plant metal extraction from contaminated soils. The aim of this study was to investigate the alteration of Ni phytoextraction by Alyssum murale, a Ni hyperaccumulator, following the application of cytokinins. The following parameters were investigated: Ni accumulation, plant growth, gas exchange, stomata behavior and the concentration of nonprotein thiols (glutathione, y-Glu-Cys, and phytochelatins). In a pot experiment, A. murale plants grown in a serpentine soil were treated with a mix of naturally occurring cytokinins. Results showed that Ni accumulation in plants ranged from 4000 to 7000 mg kg(-1) confirming the hyper-accumulation ability from the soil used. Cytokinin treatments produced a significant increase in plant biomass and transpiration rate whereas no significant variation in Ni accumulation or the concentration of non-protein thiols was observed. The results suggest that A. murale is a plant species sensitive to cytokinin treatment and that cytokinin treatment is potentially useful in increasing the phytoextraction capability by increasing biomass. Moreover, for first time, evidence was obtained that the Ni hyperaccumulation mechanism is independent of water flux and transpiration rate.     

Phenotypic characterization of microbes in the rhizosphere of Alyssum murale

Metal hyperaccumulator plants like Alyssum murale are used for phytoremediation of Ni contaminated soils. Soil microorganisms are known to play an important role in nutrient acquisition for plants, however, little is known about the rhizosphere microorganisms of hyperaccumulators. Fresh and dry weight, and Ni and Fe concentrations in plant shoots were higher when A. murale was grown in non-sterilized compared to sterilized soils. The analysis of microbial populations in the rhizosphere of A. murale and in bulk soils demonstrated that microbial numbers were affected by the presence of the plant. Significantly higher numbers of culturable actinomycetes, bacteria and fungi were found in the rhizosphere compared to bulk soil. A higher percent of Ni-resistant bacteria were also found in the rhizosphere compared to bulk soil. Percentage of acid producing bacteria was higher among the rhizosphere isolates compared to isolates from bulk soil. However, proportions of siderophore producing and phosphate solubilizing bacteria were not affected by the presence of the plant. We hypothesize that microbes in the rhizosphere of A. murale were capable of reducing soil pH leading to an increase in metal uptake by this hyperaccumulator.     

Seedling mortality of metal hyperaccumulator plants resulting from damping off by Pythium spp.

Seedling mortality of Alyssum serpyllifolium ssp. lusitanicum and A. murale , both nickel hyperaccumulators, was reduced by increasing concentrations of metal within plant tissues when inoculated with the fungi Pythium mamillatum or P. ultimum , both of which cause damping-off disease of seedlings. Pythium mamillatum , isolated from nickel-rich serpentine soil, was more tolerant of nickel than P. ultimum , isolated from low-metal control soil, and was more pathogenic than P. ultimum towards seedlings containing high concentrations of metal. These results support the hypothesis that metal hyperaccumulation by plants is closely linked to increased protection against disease.     

Soil moisture effects on uptake of metals by Thlaspi, Alyssum, and Berkheya

Most commonly used hyperaccumulator plants for phytoextraction of metals evolved on soils where moisture is limited throughout much of the year. As these plant species are commercialized for use, they are frequently moved from the point of evolution to locations where environmental conditions may be significantly different. Greatest among these potential differences is soil moisture. The objective of this study was therefore to determine whether these plants could grow in soils with much higher soil moisture and whether they would continue to hyperaccumulate metals as soils approach saturation. We examined extractable soil metal concentrations, plant growth, and metal accumulation for the Ni hyperaccumulators, Alyssum murale and Berkheya coddii and the Zn hyperaccumulators Thlaspi caerulescens cultivars AB300 and AB336. Non-hyperaccumulating control species for each were also examined. In general, extractable soil concentrations of Ni decreased with increasing soil moisture content. Few significant effects related to Zn extractability were observed for any of the soil moisture treatments. The biomass of all tested species was generally greater at higher soil moisture and inhibited at low soil moisture. Further, plants accumulated large amounts of metals from soil at higher soil moisture. Highest foliar concentrations of Zn or Ni were found at the two highest WHCs of 80 and 100%. These results show that hyperaccumulators grow well under conditions of high soil moisture content and that they continue to hyperaccumulate metals. Thus, growing Thlaspi, Alyssum, and Berkheya for commercial phytoextraction under nonnative conditions is appropriate and suggests that this technology may be applied to a wide and diverse range of soil types, climatic conditions, and irrigation regimes.     

镍污染土壤植物采矿技术关键过程及其研究进展

植物采矿是利用超积累植物高量吸收土壤中的重金属,并从中提取、冶炼金属产品,在修复污染土壤的同时实现金属的资源化。全世界广泛分布着自然风化的镍污染土壤,植物采矿因其重要的环境、生态及资源价值,被作为一种环境友好且具备经济效益的土壤修复技术,在此类地区具有广阔的应用前景。该植物采矿技术关键过程主要包括超积累植物镍高选择性根际环境响应、植物镍高效吸收转运以及生物质中镍高附加值资源化等过程。近30年,污染土壤中镍的植物采矿已经在美国、阿尔巴尼亚、马来西亚等多个国家进行了野外实践,取得了良好效果。然而,相关技术在我国的研究与应用仍然处于起步阶段。文中通过综述植物采矿技术的关键过程的研究进展,发现其中的瓶颈,为接下来植物采矿的科学研究和技术在全世界推广提供理论基础和技术指导。     

The Role of Metal Transport and Tolerance in Nickel Hyperaccumulation by Thlaspi goesingense Halacsy

Metal hyperaccumulators are plants that are capable of extracting metals from the soil and accumulating them to extraordinary concentrations in aboveground tissues (greater than 0.1% dry biomass Ni or Co or greater than 1% dry biomass Zn or Mn). Approximately 400 hyperaccumulator species have been identified, according to the analysis of field-collected specimens. Metal hyperaccumulators are interesting model organisms to study for the development of a phytoremediation technology, the use of plants to remove pollutant metals from soils. However, little is known about the molecular, biochemical, and physiological processes that result in the hyperaccumulator phenotype. We investigated the role of Ni tolerance and transport in Ni hyperaccumulation by Thlaspi goesingense, using plant biomass production, evapotranspiration, and protoplast viability assays, and by following short- and long-term uptake of Ni into roots and shoots. As long as both species (T. goesingense and Thlaspi arvense) were unaffected by Ni toxicity, the rates of Ni translocation from roots to shoots were the same in both the hyper- and nonaccumulator species. Our data suggest that Ni tolerance is sufficient to explain the Ni hyperaccumulator phenotype observed in hydroponically cultured T. goesingense when compared with the Ni-sensitive nonhyperaccumulator T. arvense.     

Variation in Nickel Content in the Nickel-Hyperaccumulating Shrub Psychotria douarrei (Rubiaceae) from New Caledonia1

Plants that hyperaccumulate Ni contain > 1000 ppm (dry wt.) in their tissues. Variation of Ni content within hyperaccumulating plant species is poorly explored. Using the Ni-hyperaccumulating shrub Psychotria douarrei, we documented variation of leaf Ni levels within individual shrubs, and variation with respect to plant size and leaf age. Plant size did not correlate significantly with leaf Ni content, and leaf Ni content did not correlate significantly with soil Ni content. Older leaves contained twice as much Ni as younger leaves. Older leaves also contained greater concentrations of Ca, Fe, and Cr but less K, P, and Cu. Five elements (Zn, Pb, Co, Mn, Mg) showed no significant variation due to leaf age. We also examined the effect of leaf age on epiphyll cover, finding increased epiphyll cover on the upper surface of older leaves. The dominant leafy liverwort epiphyll had a relatively high Ni content (400 ppm), suggesting that epiphylls of Ni hyperaccumulators obtain some Ni from host leaves. Individual shrubs differed in mean leaf Ni content almost two-fold (14,900-27,700 ppm). Variation among branches within individuals also ranged widely; however, this intraplant variability was not strongly correlated with the mean leaf Ni content of an individual shrub. We concluded that Ni contents in leaves of P. douarrei vary considerably due to leaf age, among individual shrubs, and among branches within a shrub.     

Cellular compartmentation of nickel in the hyperaccumulators Alyssum lesbiacum, Alyssum bertolonii and Thlaspi goesingense.

Nickel uptake and cellular compartmentation were investigated in three Ni hyperaccumulators: Alyssum bertolonii (Desv), Alyssum lesbiacum (Candargy) and Thlaspi goesingense (Hálácsy). The three species showed similar hyperaccumulation of Ni, but T. goesingense was less tolerant to Ni than the two Alyssum species. An addition of 500 mg Ni kg(-1) to a nutrient-rich growth medium significantly increased shoot biomass of all three species, suggesting that the Ni hyperaccumulators have a higher requirement for Ni than normal plants. Energy-dispersive X-ray microanalysis (EDXA) was performed on frozen-hydrated tissues of leaves (all species) and stems (Alyssum only). In all species analysed, Ni was distributed preferentially in the epidermal cells, most likely in the vacuoles, of the leaves and stems. In stems, there was a second peak of Ni in the boundary cells between the cortical parenchyma and the vascular cylinder. The non-glandular trichomes on the leaf surfaces of the two Alyssum species were highly enriched with Ca, but contained little Ni except in the base. In the leaves of T. goesingense, the large elongated epidermal cells contained more Ni than the cells of the stomatal complexes. The role of cellular compartmentation in Ni hyperaccumulation is discussed.     

Effects of selenium hyperaccumulation on plant-plant interactions: evidence for elemental allelopathy?

? Few studies have investigated plant-plant interactions involving hyperaccumulator plants. Here, we investigated the effect of selenium (Se) hyperaccumulation on neighboring plants. ? Soil and litter Se concentrations were determined around the hyperaccumulators Astragalus bisulcatus and Stanleya pinnata and around the nonhyperaccumulators Medicago sativa and Helianthus pumilus. We also compared surrounding vegetative cover, species composition and Se concentration in two plant species (Artemisia ludoviciana and Symphyotrichum ericoides) growing either close to or far from Se hyperaccumulators. Then, Arabidopsis thaliana germination and growth were compared on soils collected next to the hyperaccumulators and the nonhyperaccumulators. ? Soil collected around hyperaccumulators contained more Se (up to 266 mg Se kg(-1) ) than soil collected around nonhyperaccumulators. Vegetative ground cover was 10% lower around Se hyperaccumulators compared with nonhyperaccumulators. The Se concentration was higher in neighboring species A. ludoviciana and S. ericoides when growing close to, compared with far from, Se hyperaccumulators. A. thaliana showed reduced germination and growth, and higher Se accumulation, when grown on soil collected around Se hyperaccumulators compared with soil collected around nonaccumulators. ? In conclusion, Se hyperaccumulators may increase the surrounding soil Se concentration (phytoenrichment). The enhanced soil Se contents around hyperaccumulators can impair the growth of Se-sensitive plant species, pointing to a possible role of Se hyperaccumulation in elemental allelopathy.     

芦竹对不同重金属耐性的研究

研究芦竹(Arundo donax)在不同重金属污染湿地中的耐毒性能,测定了不同生长时段芦竹的生物性状和叶绿素含量,以及土壤中重金属含量的变化.结果表明,芦竹分别在浓度为100 mg·kg^-1左右的CuCu²⁺、Pb²⁺、Cd²⁺、Zn²⁺、Ni²⁺、Hg²⁺和50 mg·kg^-1以下的Cr⁶⁺污染环境中能正常成活,在40 d的生长期内,植物体内叶绿素有不同程度降低,下降比率在20%～56%,植物出现叶片软化,叶尖枯黄等症状,但植株仍呈现增长趋势.与对照植物相比较,在重金属胁迫下,植株细长,茎、叶呈黄绿色,除Cr⁶⁺、Hg²⁺外,植物高度基本不受重金属胁迫的影响.芦竹在高浓度(100mg·kg^-1)Cr⁶⁺污染环境中耐性较弱,表现出生长缓慢,部分地下茎腐烂,叶片短时间内出现枯萎等症状.结果还表明,土壤中重金属浓度随植物生长期增长而降低,除被植物吸收,植物挥发外,还存在着重金属向根际圈环境迁移的趋势,根周边湿土中重金属含量,明显高于试验缸外围湿土中重金属含量.可以认为,芦竹具有生物量大,根系发达,适应性强等特点,对修复湿地重金属污染蕴藏着巨大潜力,研究芦竹在植物修复技术中的应用,具有一定的现实意义.     

Species adaptation in serpentine soils in Lesbos Island (Greece): metal hyperaccumulation and tolerance

Serpentine (ultramafic) soils, containing relatively high nickel and other metal concentrations, present a stressful environment for plant growth but also a preferred substrate for some plants which accumulate nickel in their tissues. In the present study we focused on: (1) the relationships between serpentine soils of Lesbos Island (Greece) and serpentinophilic species in order to test their adaptation to the ‘serpentine syndrome’, and (2) the Ni-hyperaccumulation capacity of Alyssum lesbiacum, a serpentine endemic, Ni-hyperaccumulating species, recorded over all its distribution for the first time. We sampled soil and the most abundant plant species from the four serpentine localities of Lesbos Island. Soil and leaf elemental concentrations were measured across all the sites. Our results confirmed our hypothesis that serpentinophilic species are adapted to elevated heavy metal soil concentrations but restricting heavy metal concentration in their leaves. We demonstrated that different A. lesbiacum populations from Lesbos Island present differences in Ni hyperaccumulation according to soil Ni availability. Our results highlighted the understanding of serpentine ecosystems through an extensive field study in an unexplored area. Alyssum lesbiacum and Thlaspi ochroleucum emerge as two strong Ni hyperaccumulators with the former having a high potential for phytoextraction purposes.     

Accumulation and distribution of heavy metals in a simulated mangrove system treated with sewage

Constructed tide tanks were used to examine the accumulation and distribution of heavy metals in various components of a simulated mangrove ecosystem. Young Kandelia candel plants grown in mangrove soils were irrigated with wastewater of various strengths twice a week for a period of one year. The amounts of heavy metals released via tidal water and leaf litter were monitored at regular time intervals. The quantities of heavy metals retained in mangrove soil and various plant parts were also determined. Results show that most heavy metals from wastewater were retained in soils with little being uptake by plants or released into tidal seawater. However, the amounts of metals retained in plants on a per unit dry weight base were higher than those in soils as the biomass production from the young mangrove plants was much smaller when compared to the vast quantity of soils used in this study. A significantly higher heavy metal content was found in roots than in the aerial parts of the mangrove plant,indicating that the roots act as a barrier for metal translocation and protect the sensitive parts of the plant from metal contamination. In both soil and plant, concentrations of Zn, Cd, Pb and Ni increased with the strengths of wastewater, although the bioaccumulation factors for these metals decreased when wastewater strengths increased. These results suggest that the mangrove soil component has a large capacity to retain heavy metals, and the role of mangrove plants in retaining metals will depend on plant age and their biomass production. This revised version was published online in August 2006 with corrections to the Cover Date.     

HEAVY METAL ACCUMULATION AND THE NATURE OF EDAPHIC ENDEMISM IN THE GENUS CALOCHORTUS (LILIACEAE)

Bulb and leaf tissues from five species in the genus Calochortus and the soils from collection sites were analyzed for cation and trace element concentrations. All five species, only three of which are considered ultramafic endemics, possessed high concentrations of nickel and copper in both the bulbs and leaves, but not at levels high enough (> 1% dry wt) to be considered hyperaccumulators. Only moderate to trace amounts of cobalt and chromium were detected in both plant tissues. Calcium-magnesium ratios, typically low in ultramafic soils and plants, were low in bulb tissue, but two to 10 times higher in leaf tissue. The ability to tolerate excessively high levels of nickel and other heavy metals may be a physiological exaptation of the genus Calochortus and not necessarily an evolutionary response by several species to life on an ultramafic substrate. The nature of “serpentine” endemism is discussed in the context of heavy metal accumulation by various plant species.     

Performance of rose scented geranium (Pelargonium graveolens) in heavy metal polluted soil vis-à-vis phytoaccumulation of metals

An investigation was carried out to evaluate the effect of heavy metal toxicity on growth, herb, oil yield and quality and metal accumulation in rose scented geranium (Pelargonium graveolens) grown in heavy metal enriched soils. Four heavy metals (Cd, Ni, Cr, and Pb) each at two levels (10 and 20 mg kg–1 soil) were tested on geranium. Results indicated that Cr concentration in soil at 20 mg kg–1 reduced leaves, stem and root yield by 70, 83, and 45%, respectively, over control. Root growth was significantly affected in Cr stressed soil. Nickel, Cr, and Cd concentration and accumulation in plant increased with higher application of these metals. Chromium, nickel and cadmium uptake was observed to be higher in leaves than in stem and roots. Essential oil constituents were generally not significantly affected by heavy metals except Pb at 10 and 20 ppm, which significantly increased the content of citronellol and Ni at 20 ppm increased the content of geraniol. Looking in to the higher accumulation of toxic metals by geranium and the minimal impact of heavy metals on quality of essential oil, geranium can be commercially cultivated in heavy metal polluted soil for production of high value essential oil.     

Plant and rhizosphere processes involved in phytoremediation of metal-contaminated soils

This paper reviews the recent advances in understanding of metal removal from contaminated soils, using either hyperaccumulator plants, or high biomass crop species after soil treatment with chelating compounds. Progress has been made at the physiology and molecular level regarding Zn and Ni uptake and translocation in some hyperaccumulators. It is also known that natural hyperaccumulators do not use rhizosphere acidification to enhance their metal uptake. Recently, it has been found that some natural hyperaccumulators proliferate their roots positively in patches of high metal availability. In contrast, non-accumulators actively avoid these areas, and this is one of the mechanisms by which hyperaccumulators absorb more metals when grown in the same soil. However, there are few studies on the exudation and persistence of natural chelating compounds by these plants. It is thought that rhizosphere microorganisms are not important for the hyperaccumulation of metals from soil. Applications of chelates have been shown to induce large accumulations of metals like Pb, U and Au in the shoots of non-hyperaccumulators, by increasing metal solubility and root to shoot translocation. The efficiency of metal uptake does vary with soil properties, and a full understanding of the relative importance of mass flow and diffusion in the presence and absence of artificial chelates is not available. To successfully manipulate and optimise future phytoextraction technologies, it is argued that a fully combined understanding of soil supply and plant uptake is needed.